Effect Of Parathyroid Hormone On The Adipogenic Potential Of Rat Bone Marrow Mesenchymal Stem Cells

Objective:Isolation,culture and characterize of rat BMSCs,The morphology of cells were observed and cell surface marker were examined.Observing the differentiation of bone marrow mesenchymal stem cells into adipocytes and osteoblasts.Observing effect of different concentrations of parathyroid hormone on the adipogenic potential of rat bone marrow mesenchymal stem cells and explore its possible mechanism.Methods:1.Rat bone marrow mesenchymal cells were separated and expanded by adherent culture.The morphology of cells and cell growth curves was observed,cell surface marker and the cell cycle were examined by flow cytometry.2.The multilineage differentiation capability of cells was examined by culturing cells under conditions favorable for adipogenic and osteogenic differentiation.3.Taken P3 of BMSCs for test, different concentrations of PTH1-34(0, 10-10, 10-9,10-8mol/L) are respectively stimulated BMSCs, after 14 days lipoprotein lipase(LPL)activity were measured by ELISA,m RNA expression of ALP and PPARγ-2 were measured by Realtime PCR.4.Statistical analysis:data are presented as X ±S,All statistical analysis is performed with windows SPSS 13.0.One-way ANOVE can determine the different between the group.LSD determine the different between random two groups.Differences are considered significant at a value of P<0.05.Results:1.BMSCs adherent growth, uniform long shuttle shape, growth curve for the "S" type.The cell cycle was mostly in G1/G0 phase.The cells expressed the stemness markers CD44 and CD29 and negative expressed CD45.2.Oil Red O staining BMSCs after three weeks of culture demonstrated numerous intracellular lipid droplets. Alizarin red staining BMSCs after four weeks of culturedemonstrated red calcium nodules.3.Compared with the control group,different concentrations of PTH1-34 groups decreased m RNA expression of ALP and PPARγ-2,LPL activity were increased.Conclusions:1.BMSCs can be purification and amplification by using adherent culture method,expression of CD44 and CD29,no expression of CD45.BMSCs showed the osteogenic and adipogenic different iation potential.2.PTH1-34 inhibited BMSCs adipogenic differentiation, promoted osteogenic differentiation, and showed dose dependent.3.PTH1-34 may be inhibit BMSCs differentiation into adipocytes by down-regulation of PPARγ-2.